马疱疹病毒gD蛋白主要抗原域的表达及初步应用  

Prokaryotic expression and preliminary application of equine herpes virus gD protein

作　　者：时成龙[1,2] 侯玉杰[1,2] 相文华[2] 郭巍[2] 李红梅[2] 赵立平[2] 何剑斌[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110161 [2]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001

出　　处：《中国兽医杂志》2010年第6期12-15,共4页Chinese Journal of Veterinary Medicine

摘　　要：根据GenBank马疱疹病毒1型(EHV-1)和4型(EHV-4)gD基因进行分析,选取同源性较高且抗原性强的C端序列设计一对引物进行扩增。将扩增的基因插入pET-30a的BamHⅠ和SalⅠ之间构建原核表达载体pET-gD。然后将pET-gD质粒转化至BL21(DE3)宿主菌,对培养和表达条件进行优化,实现EHV gD蛋白主要抗原区域的高效表达。将重组pET-gD蛋白进行纯化并接种长白兔制备高免血清。经免疫印迹试验鉴定,证实所得纯化表达产物具有良好的抗原性。According to GenBank,a pair of primers were designed to amplify equine herpes virus type 1 and 4 gD gene.The amplified gene was inserted into pET30a between BamHⅠ and SaⅡ,built prokaryotic expression vector pET-gD1.Then pET-gD1 plasmid was transformed into BL 21（DE3） host bacteria,the EHV gD protein major antigen domain was highly expressed.The recombinant protein was purified and prepare high-titer-immuno-serum of Changbai rabbits.Western blot result indicated that the purified protein product has a good antigenicity.

关 键 词：马疱疹病毒 gD蛋白 原核表达 免疫原性 

分 类 号：S852.659.1[农业科学—基础兽医学]